Sample positioning method and system

Abstract

The invention provides a sample positioning method and system, and the method comprises the steps: obtaining a first target point coordinate of a sample target point and a coordinate of at least one mark point under a first microscopic imaging system, and obtaining a corresponding coordinate of the mark point under a second microscopic imaging system; calculating a coordinate conversion relation between the first microscopic imaging system and the second microscopic imaging system, converting the first target point coordinate into a second target point coordinate under the second microscopic imaging system by using the coordinate conversion relation, and adjusting the position of the sample in the second microscopic imaging system according to the second target point coordinate. Thus, by automatically collecting and calculating the coordinate conversion relation, accurate alignment of sample target points among different microscopic imaging systems can be efficiently achieved, errors caused by traditional manual alignment are avoided, experiment repeatability and data accuracy are improved, operation efficiency is improved, and the method is suitable for various microscopic imaging systems.

Description

Technical Field

[0001] The present invention relates to the field of optical technology, and particularly to a sample positioning method and system thereof. Background Art

[0002] The multi-system coupling technology can significantly improve the analysis ability by combining different microscopic imaging systems (such as optical microscopes and scanning electron microscopes) to obtain multi-dimensional information of samples.

[0003] However, when switching between different microscopic imaging systems, the position of the sample may change due to factors such as system calibration differences, vibration, or thermal expansion, resulting in a decrease in positioning accuracy and affecting the accuracy of subsequent analysis. The position of the target point will shift, making it difficult to accurately reproduce the initial observation position.

[0004] The existing sample positioning methods usually adopt the following technical means:

[0005] Manual alignment: Rely on the operator to manually adjust the position of the sample until the target point is found in the second microscopic imaging system. This method is time-consuming and has large errors, and is difficult to apply to high-precision experiments.

[0006] Mechanical alignment: Rely on high-precision mechanical devices to move and rotate the sample stage for adjustment, but the equipment is expensive and it is difficult to meet the coordinate conversion requirements between different microscopic imaging systems.

[0007] Therefore, the sample positioning accuracy of the related technology is poor and the efficiency is low.

[0008] It should be noted that the information disclosed in the above background art section is only used to enhance the understanding of the background of the present invention, and thus may include information that does not constitute the prior art known to those of ordinary skill in the art. Summary of the Invention

[0009] Aiming at the problems in the prior art, the purpose of the present invention is to provide a sample positioning method and system thereof, which overcome the difficulties of the prior art and can solve the technical problems of poor sample positioning accuracy and low efficiency in the related technology.

[0010] An embodiment of the present disclosure provides a sample positioning method, which is applicable to sample positioning across a first microscopic imaging system and a second microscopic imaging system. The sample positioning method includes:

[0011] Under the first microscopic imaging system, obtain the first target point coordinates of the sample target point and the coordinates of at least one marker point, where the marker point includes a base marker point and / or a sample marker point;

[0012] Obtain the corresponding coordinates of the marker point under the second microscopic imaging system;

[0013] Calculate the coordinate transformation relationship between the first microscopic imaging system and the second microscopic imaging system based on the coordinates of the marked points under the first microscopic imaging system and the second microscopic imaging system, and use the coordinate transformation relationship to convert the first target point coordinates into the second target point coordinates under the second microscopic imaging system;

[0014] Adjust the position of the sample in the second microscopic imaging system according to the second target point coordinates.

[0015] Optionally, the first target point coordinates and the coordinates of at least one marked point are both two-dimensional coordinates;

[0016] The marked points include the base marked points of the sample base, the coordinates of the base marked points under the first microscopic imaging system are the first base marked point coordinates, and the coordinates under the second microscopic imaging system are the second base marked point coordinates;

[0017] Adopt the following method to obtain the rotation angle of the sample base as the coordinate transformation relationship:

[0018] Select a base reference point on the sample base, and obtain its first reference point coordinates under the first microscopic imaging system and the second reference point coordinates under the second microscopic imaging system;

[0019] Based on the first relative coordinates between the first base marked point coordinates and the first reference point coordinates, and the second relative coordinates between the second base marked point coordinates and the second reference point coordinates, calculate the rotation angle of the sample base, so as to use the rotation angle of the sample base to convert the first target point coordinates into the second target point coordinates.

[0020] Optionally, the step of using the rotation angle of the sample base to convert the first target point coordinates into the second target point coordinates includes:

[0021] Calculate the third relative coordinates between the first target point coordinates and the first reference point coordinates, use the rotation angle of the sample base to convert the third relative coordinates into the fourth relative coordinates under the second microscopic imaging system, and calculate the second target point coordinates according to the fourth relative coordinates and the second reference point coordinates.

[0022] Optionally, the marked points include the base marked points and the sample marked points, and the coordinates of the sample marked points under the first microscopic imaging system are the first marked point coordinates;

[0023] The step of using the rotation angle of the sample base to convert the first target point coordinates into the second target point coordinates includes:

[0024] Calculate a fifth relative coordinate between the coordinates of the first marked point and the coordinates of the first reference point, convert the fifth relative coordinate into a sixth relative coordinate under the second microscopic imaging system by using the rotation angle of the sample base, and calculate the coordinates of the second marked point of the sample marked point according to the sixth relative coordinate and the coordinates of the second reference point;

[0025] Calculate a seventh relative coordinate between the coordinates of the first target point and the coordinates of the first marked point, convert the seventh relative coordinate into an eighth relative coordinate under the second microscopic imaging system by using the rotation angle of the sample base, and calculate the coordinates of the second target point according to the eighth relative coordinate and the coordinates of the second marked point.

[0026] Optionally, the marked points include the base marked point and the sample marked point, and the coordinates of the sample marked point under the first microscopic imaging system are the coordinates of the first marked point;

[0027] The conversion of the coordinates of the first target point into the coordinates of the second target point by using the rotation angle of the sample base includes:

[0028] Calculate a fifth relative coordinate between the coordinates of the first marked point and the coordinates of the first reference point, convert the fifth relative coordinate into a sixth relative coordinate under the second microscopic imaging system by using the rotation angle of the sample base, and calculate the coordinates of the second marked point of the sample marked point according to the sixth relative coordinate and the coordinates of the second reference point;

[0029] Select a sample reference point on the sample surface of the sample, and obtain its first sample reference point coordinates under the first microscopic imaging system and its second sample reference point coordinates under the second microscopic imaging system;

[0030] Based on the ninth relative coordinate between the coordinates of the first marked point and the first sample reference point, and the tenth relative coordinate between the coordinates of the second marked point and the second sample reference point, calculate the rotation angle of the sample surface;

[0031] Calculate an eleventh relative coordinate between the coordinates of the first target point and the first sample reference point, convert the eleventh relative coordinate into a twelfth relative coordinate under the second microscopic imaging system by using the rotation angle of the sample surface, and calculate the coordinates of the second target point according to the twelfth relative coordinate and the coordinates of the second sample reference point.

[0032] Optionally, there are multiple base marked points; the base reference point is one of the multiple base marked points, or the base reference point is any point on the base outside the multiple base marked points.

[0033] Optionally, the coordinates of the first target point and the coordinates of at least one marked point are all three-dimensional coordinates;

[0034] The marked points include the base marked points of the sample base, the coordinates of the base marked points under the first microscopic imaging system are the first base marked point coordinates, and the coordinates under the second microscopic imaging system are the second base marked point coordinates;

[0035] The following method is used to obtain the rotation matrix of the sample base as the coordinate transformation relationship:

[0036] Select a base reference point on the sample base, and obtain its first reference point coordinates under the first microscopic imaging system and its second reference point coordinates under the second microscopic imaging system;

[0037] Calculate a first vector of the first base marked point coordinates relative to the first reference point coordinates, and calculate a second vector of the second base marked point coordinates relative to the second reference point coordinates;

[0038] Select a first group of first vectors and a first group of second vectors. The first group of first vectors includes the first vectors of two linearly independent base marked points, and the first group of second vectors includes the second vectors of the two linearly independent base marked points. Calculate a first unit normal vector based on the first group of first vectors, and calculate a second unit normal vector based on the first group of second vectors;

[0039] Select a second group of first vectors that are linearly independent of the first unit normal vector and a second group of second vectors that are linearly independent of the second unit normal vector. Normalize the second group of first vectors to obtain a first group of unit vectors, and normalize the second group of second vectors to obtain a second group of unit vectors. The first group of unit vectors includes two first unit vectors, and the second group of unit vectors includes two second unit vectors;

[0040] Perform a cross product calculation of a third unit vector based on the first first unit vector in the first group of unit vectors and the first unit normal vector, and perform a cross product calculation of a fourth unit vector based on the first second unit vector in the second group of unit vectors and the second unit normal vector;

[0041] Construct a first basis vector matrix from the second first unit vector, the first unit normal vector, and the third unit vector in the first group of unit vectors, and construct a second basis vector matrix from the second unit vector, the second unit normal vector, and the fourth unit vector in the second group of unit vectors;

[0042] Calculate the rotation matrix of the sample base, which is obtained by multiplying the first basis vector matrix by the transpose matrix of the second basis vector matrix, so as to convert the first target point coordinates into the second target point coordinates by using the rotation matrix of the sample base.

[0043] Optionally, converting the first target point coordinates into the second target point coordinates by using the rotation matrix of the sample base includes:

[0044] Calculate the third vector of the first target point coordinates relative to the first reference point coordinates, and multiply the third vector by the first basis vector matrix to obtain the first basis vector representation;

[0045] Multiply the rotation matrix of the sample base by the first basis vector representation to obtain the fourth vector under the second microscopic imaging system;

[0046] Use the second basis vector matrix to convert the fourth vector into the second basis vector representation, and add the second basis vector representation to the second reference point coordinates to obtain the second target point coordinates.

[0047] Optionally, the marking points include the base marking points and the sample marking points, and the coordinates of the sample marking points under the first microscopic imaging system are the first marking point coordinates;

[0048] The converting the first target point coordinates into the second target point coordinates by using the rotation matrix of the sample base includes:

[0049] Calculate the fifth vector of the first marking point coordinates relative to the first reference point coordinates, multiply the fifth vector by the first basis vector matrix to obtain the third basis vector representation, multiply the rotation matrix of the sample base by the third basis vector representation to obtain the sixth vector under the second microscopic imaging system, use the second basis vector matrix to convert the sixth vector into the fourth basis vector representation, and add the second basis vector representation to the second reference point coordinates to obtain the second marking point coordinates under the second microscopic imaging system;

[0050] Calculate the seventh vector of the first target point coordinates relative to the first marking point coordinates, multiply the seventh vector by the first basis vector matrix to obtain the fifth basis vector representation, multiply the rotation matrix of the sample base by the fifth basis vector representation to obtain the eighth vector under the second microscopic imaging system, use the second basis vector matrix to convert the eighth vector into the sixth basis vector representation, and add the second basis vector representation to the second marking point coordinates to obtain the second target point coordinates.

[0051] Optionally, the marked points include the base marked points and the sample marked points, and the coordinates of the sample marked points under the first microscopic imaging system are the first marked point coordinates;

[0052] The conversion of the first target point coordinates into the second target point coordinates by using the rotation matrix of the sample base includes:

[0053] Calculating a ninth vector of the first marked point coordinates relative to the first reference point coordinates, multiplying the ninth vector by the first basis vector matrix to obtain a fifth basis vector representation, multiplying the rotation matrix of the sample base by the fifth basis vector representation to obtain a tenth vector under the second microscopic imaging system, converting the tenth vector into a sixth basis vector representation by using the second basis vector matrix, and adding the sixth basis vector representation to the second reference point coordinates to obtain the second marked point coordinates under the second microscopic imaging system;

[0054] Selecting a sample reference point on the sample surface of the sample, and obtaining its first sample reference point coordinates under the first microscopic imaging system and its second sample reference point coordinates under the second microscopic imaging system;

[0055] Calculating an eleventh vector of the first sample marked point coordinates relative to the first sample reference point coordinates, and calculating a twelfth vector of the second base marked point coordinates relative to the second reference point coordinates;

[0056] Selecting a first group of eleventh vectors and a first group of twelfth vectors, the first group of eleventh vectors including the eleventh vectors of two linearly independent base marked points, the first group of twelfth vectors including the twelfth vectors corresponding to the same base marked points as the eleventh vectors, calculating a third unit normal vector based on the first group of eleventh vectors, and calculating a fourth unit normal vector based on the first group of twelfth vectors;

[0057] Selecting a second group of eleventh vectors that are linearly independent of the third unit normal vector and a second group of twelfth vectors that are linearly independent of the fourth unit normal vector and correspond to the same sample marked points as the second group of eleventh vectors, normalizing the second group of eleventh vectors to obtain a third group of unit vectors, and normalizing the second group of twelfth vectors to obtain a fourth group of unit vectors, the third group of unit vectors including two fifth unit vectors, and the fourth group of unit vectors including two sixth unit vectors;

[0058] Calculating a seventh unit vector by performing a cross product of the first fifth unit vector in the third group of unit vectors and the third unit normal vector, and calculating an eighth unit vector by performing a cross product of the first sixth unit vector in the fourth group of unit vectors and the fourth unit normal vector;

[0059] Construct a third basis vector matrix from the second fifth unit vector, the third unit normal vector, and the seventh unit vector in the third group of unit vectors, and construct a fourth basis vector matrix from the sixth unit vector, the fourth unit normal vector, and the eighth unit vector in the fourth group of unit vectors;

[0060] Calculate the rotation matrix of the sample plane, which is obtained by multiplying the third basis vector matrix by the transpose matrix of the fourth basis vector matrix;

[0061] Calculate the thirteenth vector of the first target point coordinate relative to the first sample reference point coordinate, multiply the thirteenth vector by the third basis vector matrix to obtain a seventh basis vector representation, multiply the seventh basis vector representation by the rotation matrix of the sample plane to obtain a fourteenth vector under the second microscopic imaging system, convert the fourteenth vector into an eighth basis vector representation using the fourth basis vector matrix, and add the eighth basis vector representation to the second sample reference point coordinate to obtain the second target point coordinate.

[0062] A second aspect of the present disclosure provides a sample positioning system, which is applicable to sample positioning across a first microscopic imaging system and a second microscopic imaging system. The sample positioning system includes:

[0063] A first acquisition module, under the first microscopic imaging system, acquires the first target point coordinate of the sample target point and the coordinates of at least one marker point, where the marker point includes a base marker point and / or a sample marker point;

[0064] A second acquisition module, acquires the coordinates corresponding to the marker point under the second microscopic imaging system;

[0065] A calculation module, based on the coordinates of the marker point under the first microscopic imaging system and the second microscopic imaging system, calculates the coordinate conversion relationship between the first microscopic imaging system and the second microscopic imaging system, and uses the coordinate conversion relationship to convert the first target point coordinate into the second target point coordinate under the second microscopic imaging system;

[0066] An adjustment module, adjusts the position of the sample in the second microscopic imaging system according to the second target point coordinate.

[0067] The sample positioning method and system proposed by the embodiments of the present disclosure have the following advantages:

[0068] Under the first microscopic imaging system, the first target point coordinates of the sample target point and the coordinates of at least one marker point are obtained, the corresponding coordinates of the marker point under the second microscopic imaging system are obtained, and based on the coordinates of the marker point under the first microscopic imaging system and the second microscopic imaging system, the coordinate conversion relationship between the first microscopic imaging system and the second microscopic imaging system is calculated, and the first target point coordinates are converted into the second target point coordinates under the second microscopic imaging system by using the coordinate conversion relationship. According to the second target point coordinates, the position of the sample in the second microscopic imaging system is adjusted. In this way, by automatically collecting and calculating the coordinate conversion relationship, it has good anti-interference ability against the small changes in the position of the sample caused by physical properties (such as vibration, thermal expansion, etc.), and at the same time has good processing effects on the physical position changes such as displacement, rotation, and tilt generated by the sample placement. It can efficiently achieve the precise alignment of the sample target points between different microscopic imaging systems, avoid the errors introduced by traditional manual alignment, improve the experimental repeatability and data accuracy, improve the operation efficiency, and is applicable to a variety of microscopic imaging systems.

[0069] It should be understood that the above general description and the following detailed description are only exemplary and explanatory, and cannot limit the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

[0070] Other features, objects, and advantages of the present invention will become more apparent by reading the following detailed description of non-limiting embodiments with reference to the accompanying drawings.

[0071] Figure 1 It is a flowchart of the sample positioning method provided by the embodiment of the present disclosure;

[0072] Figure 2 For applicability Figure 1 It is a schematic scene diagram of a microscopic imaging system for the sample positioning method shown;

[0073] Figure 3 It is one of the flowcharts of the sample positioning method provided by the embodiment of the present disclosure in a two-dimensional positioning scenario;

[0074] Figure 4 For Figure 3 It is a schematic principle diagram of the coordinate system of the first microscopic imaging system and the coordinate system of the second microscopic imaging system in the sample positioning method shown;

[0075] Figure 5 It is the second of the flowcharts of the sample positioning method provided by the embodiment of the present disclosure in a two-dimensional positioning scenario;

[0076] Figure 6 For Figure 5 It is a schematic principle diagram of the coordinate system of the first microscopic imaging system and the coordinate system of the second microscopic imaging system in the sample positioning method shown;

[0077] Figure 7 The third flowchart of the sample positioning method provided by the present disclosure in a two-dimensional positioning scenario;

[0078] Figure 8 For Figure 7 Schematic diagram of the principle of the first microscopic imaging system coordinate system and the second microscopic imaging system coordinate system in the sample positioning method shown;

[0079] Figure 9 The first flowchart of the sample positioning method provided by the present disclosure in a three-dimensional positioning scenario;

[0080] Figure 10 The second flowchart of the sample positioning method provided by the present disclosure in a three-dimensional positioning scenario;

[0081] Figure 11 The third flowchart of the sample positioning method provided by the present disclosure in a three-dimensional positioning scenario;

[0082] Figure 12 The fourth flowchart of the sample positioning method provided by the present disclosure in a three-dimensional positioning scenario;

[0083] Figure 13 Shows a schematic diagram of a module structure of the sample positioning system provided by the present disclosure. Detailed implementation manners

[0084] Now, example embodiments will be described more fully with reference to the accompanying drawings. However, the example embodiments can be implemented in various forms and should not be construed as limited to the examples set forth herein; rather, these embodiments are provided so that this disclosure will be more complete and comprehensive, and will fully convey the concept of the example embodiments to those skilled in the art. The described features, structures, or characteristics can be combined in any suitable manner in one or more embodiments.

[0085] In addition, the accompanying drawings are only schematic illustrations of the present disclosure and are not necessarily drawn to scale. The same reference numerals in the drawings denote the same or similar parts, and thus repeated descriptions thereof will be omitted. Some of the block diagrams shown in the drawings are functional entities and do not necessarily correspond to physically or logically independent entities. These functional entities can be implemented in software form, or in one or more hardware modules or integrated circuits, or in different networks and / or processor devices and / or microcontroller devices.

[0086] In addition, the concepts such as "first" and "second" mentioned in the present disclosure are only used to distinguish different devices, modules, or units, and are not used to limit the order of the functions performed by these devices, modules, or units or their interdependent relationships.

[0087] The embodiments of the present disclosure provide an improved sample positioning method. By calculating the coordinate transformation relationship of the marking points, high-precision conversion of the sample target points across microscopic imaging systems is achieved, improving the positioning accuracy and automation level.

[0088] Exemplarily, taking the transfer of a sample between a first microscopic imaging system and a second microscopic imaging system as an example, this method aims to achieve precise position conversion of the sample between the first microscopic imaging system and the second microscopic imaging system. This method is particularly applicable to scenarios that require cross-system analysis, such as maintaining the position consistency of the target points when transferring samples between different microscopes.

[0089] As Figure 1 shown, the sample positioning method includes but is not limited to the following steps:

[0090] Step 110: Under the first microscopic imaging system, obtain the first target point coordinates of the sample target point and the coordinates of at least one marking point, where the marking point includes a base marking point and / or a sample marking point;

[0091] Step 120: Obtain the corresponding coordinates of the marking point under the second microscopic imaging system;

[0092] Step 130: According to the coordinates of the marking point under the first microscopic imaging system and the second microscopic imaging system, calculate the coordinate transformation relationship between the first microscopic imaging system and the second microscopic imaging system, and use the coordinate transformation relationship to convert the first target point coordinates into the second target point coordinates under the second microscopic imaging system;

[0093] Step 140: Adjust the position of the sample in the second microscopic imaging system according to the second target point coordinates.

[0094] In the embodiments of the present disclosure, the microscopic imaging system refers to imaging devices that can obtain sample images, including but not limited to optical microscopes, electron microscopes, confocal microscopes, etc.

[0095] Combined with Figure 2 shown, a nail table 2 is provided on the sample base 1, and the sample 3 is placed on the nail table 2. The marking point refers to a reference point set on the sample 3 or the sample base 1 that can be used for coordinate matching, including a base marking point 4 and a sample marking point 5. The sample target point 6 refers to the sample area that needs to maintain a consistent position under different microscopic systems during the experiment.

[0096] Among them, the coordinate transformation relationship: refers to the mathematical mapping relationship between the first microscopic imaging system and the second microscopic imaging system, including a two-dimensional rotation angle or a three-dimensional rotation matrix, which will be described in detail later.

[0097] In this embodiment, first, under the first microscopic imaging system (such as an optical microscope), the coordinates of the target point of the sample (denoted as the first target point coordinate P A6 ) and a set of marker point coordinates are automatically or semi-automatically obtained through image processing software. These marker points include the "base marker points" set on the sample base and (optionally) the "sample marker points" set on the sample body. Subsequently, in the second microscopic imaging system (such as an electron microscope or a confocal microscope), the marker point coordinates (denoted as P B1 , P B2 , ……) of the same physical sample are repeatedly collected to ensure a one-to-one correspondence of the marker points under the two systems.

[0098] Using the marker point coordinates under the two microscopic imaging systems, the coordinate transformation relationship between the two microscopic imaging systems can be calculated according to the corresponding algorithm (two-dimensional or three-dimensional), that is, the target point coordinate P A6 under the first microscopic imaging system is converted into the target point coordinate P B6 under the second microscopic imaging system. After the calculation is completed, the position of the sample in the second microscopic imaging system is adjusted according to P B6 to achieve in-situ positioning.

[0099] Therefore, this embodiment has good anti-interference ability against the small changes in the position of the sample caused by physical properties (such as vibration, thermal expansion, etc.) by automatically collecting and calculating the coordinate transformation relationship. At the same time, it has a good processing effect on the physical position changes such as displacement, rotation, and tilt generated by the placement of the sample, can efficiently achieve the precise alignment of the sample target points between different microscopic imaging systems, avoid the errors introduced by traditional manual alignment, improve the experimental repeatability and data accuracy, improve the operation efficiency, and is applicable to a variety of microscopic imaging systems.

[0100] The following will specifically elaborate on the above Figure 1 sample positioning method.

[0101] In the two-dimensional positioning scenario, the first target point coordinate and the coordinates of at least one marker point are all two-dimensional coordinates. The marker points include the base marker points of the sample base. The coordinates of the base marker points under the first microscopic imaging system are the first base marker point coordinates, and the coordinates under the second microscopic imaging system are the second base marker point coordinates.

[0102] In this case, as Figure 3 shown, the following method is used to obtain the rotation angle of the sample base as the coordinate transformation relationship:

[0103] Step 310: Select a base reference point on the sample base and obtain its first reference point coordinates under the first microscopic imaging system and its second reference point coordinates under the second microscopic imaging system;

[0104] Step 320: Calculate the rotation angle of the sample base based on the first relative coordinates between the first base marker point coordinates and the first reference point coordinates, and the second relative coordinates between the second base marker point coordinates and the second reference point coordinates, so as to convert the first target point coordinates into the second target point coordinates by using the rotation angle of the sample base.

[0105] Specifically, in combination with Figure 4 , under the first microscopic imaging system (such as an optical microscope), obtain the first target point coordinates (x1, y1) of the sample target point and the first base marker point coordinates (x2, y2) of the base marker point 1. The base marker point is a fixed point preset on the sample base for positioning the base position.

[0106] Transfer the sample to the second microscopic imaging system (such as a scanning electron microscope), and obtain the corresponding coordinates of the same base marker point 1 under the second microscopic imaging system, that is, the second base marker point coordinates (x3, y3).

[0107] Select the base reference point 0 (which can be one of multiple base marker points or other points on the sample base other than the base marker points). For the same base reference point 0, its first reference point coordinates under the first microscopic imaging system are (x5, y5), and its second reference point coordinates under the second microscopic imaging system are (x6, y6);

[0108] Calculate the first relative coordinates (Δx1, Δy1) = (x2 - x5, y2 - y5), and convert them into polar coordinates (r1, θ1);

[0109] Calculate the second relative coordinates (Δx2, Δy2) = (x3 - x6, y3 - y6), and convert them into polar coordinates (r2, θ2).

[0110] Then, the rotation angle θ of the sample base is θ = θ2 - θ1, which reflects the rotation difference of the base in the plane.

[0111] Use the rotation angle θ to convert the first target point coordinates (x1, y1) into the second target point coordinates (x4, y4).

[0112] In one implementation, converting the first target point coordinates into the second target point coordinates by using the rotation angle of the sample base includes:

[0113] Calculate the third relative coordinates between the first target point coordinates and the first reference point coordinates, and convert the third relative coordinates into the fourth relative coordinates under the second microscopic imaging system by using the rotation angle of the sample base, and calculate the second target point coordinates according to the fourth relative coordinates and the second reference point coordinates.

[0114] Specifically, the coordinates of the first target point are (x1, y1). Calculate the third relative coordinates (Δx3, Δy3) = (x1 - x5, y1 - y5), and convert them into polar coordinates (r3, φ).

[0115] Adjust the angle to φ + θ using the rotation angle θ to obtain the fourth relative coordinates (Δx4, Δy4).

[0116] The coordinates of the second target point are (x4, y4) = (x6 + Δx4, y6 + Δy4).

[0117] This embodiment details the implementation method of two-dimensional coordinate transformation. Calculate the relative coordinates through the base marking points and the base reference points, determine the rotation angle θ of the sample base, and use it as the coordinate transformation relationship. The rotation angle reflects the planar rotation difference of the sample base between the two microscopic imaging systems. Map the target point coordinates from the first microscopic imaging system to the second microscopic imaging system through polar coordinate transformation. The base reference point is used as a benchmark to ensure the consistency of relative coordinate calculation. The above calculation method is applicable to two-dimensional positioning scenarios with elementary or medium precision. The method has high calculation efficiency and is suitable for rapid positioning requirements.

[0118] In another embodiment, the marking points include the base marking points and the sample marking points, and the coordinates of the sample marking points under the first microscopic imaging system are the first marking point coordinates.

[0119] In this case, refer to Figure 5 , and converting the first target point coordinates into the second target point coordinates using the rotation angle of the sample base includes:

[0120] Step 510: Calculate the fifth relative coordinates between the first marking point coordinates and the first reference point coordinates, convert the fifth relative coordinates into the sixth relative coordinates under the second microscopic imaging system using the rotation angle of the sample base, and calculate the second marking point coordinates of the sample marking points according to the sixth relative coordinates and the second reference point coordinates;

[0121] Step 520: Calculate the seventh relative coordinates between the first target point coordinates and the first marking point coordinates, convert the seventh relative coordinates into the eighth relative coordinates under the second microscopic imaging system using the rotation angle of the sample base, and calculate the second target point coordinates according to the eighth relative coordinates and the second marking point coordinates.

[0122] This embodiment can introduce sample marking points on the basis of preliminary positioning to further correct the coordinates of the second target point. The sample marking points are located on the surface of the sample and are closer to the target point than the base marking points, and can capture the minute rotation or displacement of the sample itself. By calculating the coordinates of the sample marking points under the second microscopic imaging system, the positioning error of the target point is corrected to improve the accuracy.

[0123] As Figure 6 shown, the position coordinates of the sample marking point 1 in the first microscopic imaging and the second microscopic imaging are shown respectively.

[0124] The specific steps are described as follows:

[0125] The first marking point coordinates of the sample marking point are (x7, y7).

[0126] Calculate the fifth relative coordinates (Δx5, Δy5) = (x7 - x5, y7 - y5), and convert them into polar coordinates (r5, φ5).

[0127] For the polar coordinates (r5, φ5), use the rotation angle θ of the sample base to adjust the angle to φ5 + θ to obtain the sixth relative coordinates (Δx6, Δy6).

[0128] Calculate the second marking point coordinates (x8, y8) = (x6 + Δx6, y6 + Δy6).

[0129] Calculate the seventh relative coordinates (Δx7, Δy7) = (x1 - x7, y1 - y7), and convert them into polar coordinates (r7, φ7).

[0130] Use the base rotation angle θ to adjust the angle to φ7 + θ to obtain the eighth relative coordinates (Δx8, Δy8).

[0131] Calculate the optimized second target point coordinates (x9, y9) = (x8 + Δx8, y8 + Δy8).

[0132] Use the optimized second target point coordinates (x9, y9) to adjust the position of the sample.

[0133] Using this embodiment, through the correction of the sample marking points, the positioning accuracy is further improved, and the minute rotation and displacement of the sample itself can be effectively compensated, which is applicable to two-dimensional positioning scenarios requiring high accuracy.

[0134] In another embodiment of the present disclosure, as Figure 7 shown, converting the first target point coordinates into the second target point coordinates by using the rotation angle of the sample base includes:

[0135] Step 710: Calculate the fifth relative coordinate between the coordinates of the first marked point and the coordinates of the first reference point, convert the fifth relative coordinate into the sixth relative coordinate under the second microscopic imaging system by using the rotation angle of the sample base, and calculate the coordinates of the second marked point of the sample mark according to the sixth relative coordinate and the coordinates of the second reference point;

[0136] Step 720: Select a sample reference point on the sample surface of the sample, and obtain its first sample reference point coordinates under the first microscopic imaging system and its second sample reference point coordinates under the second microscopic imaging system;

[0137] Step 730: Calculate the rotation angle of the sample surface based on the ninth relative coordinate between the coordinates of the first marked point and the coordinates of the first sample reference point, and the tenth relative coordinate between the coordinates of the second marked point and the coordinates of the second sample reference point;

[0138] Step 740: Calculate the eleventh relative coordinate between the coordinates of the first target point and the coordinates of the first sample reference point, convert the eleventh relative coordinate into the twelfth relative coordinate under the second microscopic imaging system by using the rotation angle of the sample surface, and calculate the coordinates of the second target point according to the twelfth relative coordinate and the coordinates of the second sample reference point.

[0139] Combined Figure 8 As shown, in this embodiment, on the basis of the above-mentioned fine positioning, a sample surface reference point is introduced to calculate the rotation angle θ of the sample surface itself s , and further correct the coordinates of the target point. The rotation angle of the sample surface reflects the independent rotation of the sample on the base. Combined with the rotation angle of the base, it can capture the position of the target point more accurately.

[0140] Combined with the above embodiment, the specific steps are as follows:

[0141] Use the above embodiment to calculate the coordinates of the second marked point (x8, y8);

[0142] Select the sample surface reference point, and its first sample reference point coordinates are (x 10 , y 10 ), and the second sample reference point coordinates are (x 11 , y 11 ).

[0143] Calculate the ninth relative coordinate (Δx9, Δy9) = (x7 - x 10 , y7 - y 10 ), convert it to polar coordinates (r9, φ9), and calculate the tenth relative coordinate (Δx 10 , Δy 10 ) = (x8 - x 11, y8 - y 11 ), convert to polar coordinates (r 10 , φ 10 ).

[0144] Sample surface rotation angle θ s = φ 10 - φ9.

[0145] Calculate the eleventh relative coordinate (Δx 11 , Δy 11 ) = (x1 - x 10 , y1 - y 10 ), convert to polar coordinates (r 11 , φ 11 ).

[0146] Use the sample surface rotation angle θ s to adjust the angle to φ 11 + θ s , and obtain the twelfth relative coordinate (Δx 12 , Δy 12 ).

[0147] Calculate the second target point coordinates (x 12 , y 12 ) = (x 11 + Δx 12 , y 11 + Δy 12 ).

[0148] Use the second target point coordinates (x 12 , y 12 ) obtained by fine positioning to adjust the sample position.

[0149] Through sample surface rotation correction, the positioning accuracy is further improved, and it can accurately compensate for the rotation deviation of the sample itself, which is applicable to high-precision two-dimensional analysis scenarios.

[0150] In addition. There are usually multiple base marking points, and the base reference point can be one of them or other points on the base. This flexibility allows the method to adapt to different base designs and reduces the dependence on the number of marking points.

[0151] For example, if there are multiple base marking points, such as (x2, y2), (x 13 , y 13 ), select (x2, y2) as the base reference point, its first reference point coordinates are (x2, y2), and the second reference point coordinates are (x3, y3).

[0152] For example, the base reference point is any point on the base: select a non-marking point on the base and input its first reference point coordinates (x5, y5) and second reference point coordinates (x6, y6).

[0153] The flexible selection of the base reference point improves the generality of the method and is applicable to scenarios where the number of marked points is limited or unevenly distributed.

[0154] In a three-dimensional positioning scenario, the coordinates of the first target point and at least one marked point are all three-dimensional coordinates. The marked points include the base marked points of the sample base, and the coordinates of the base marked points under the first microscopic imaging system are the first base marked point coordinates, and the coordinates under the second microscopic imaging system are the second base marked point coordinates;

[0155] As Figure 9 shown, the following method is used to obtain the rotation matrix of the sample base as the coordinate transformation relationship:

[0156] Step 910: Select the base reference point on the sample base and obtain its first reference point coordinates under the first microscopic imaging system and its second reference point coordinates under the second microscopic imaging system;

[0157] Step 920: Select a first set of first vectors and a first set of second vectors. The first set of first vectors includes the first vectors of two linearly independent base marked points, and the first set of second vectors includes the second vectors of the two linearly independent base marked points. Calculate the first unit normal vector based on the first set of first vectors, and calculate the second unit normal vector based on the first set of second vectors;

[0158] Step 930: Select a second set of first vectors that are linearly independent of the first unit normal vector and a second set of second vectors that are linearly independent of the second unit normal vector. Normalize the second set of first vectors to obtain a first set of unit vectors, and normalize the second set of second vectors to obtain a second set of unit vectors. The first set of unit vectors includes two first unit vectors, and the second set of unit vectors includes two second unit vectors;

[0159] Step 940: Calculate the third unit vector by cross-multiplying the first first unit vector in the first set of unit vectors and the first unit normal vector, and calculate the fourth unit vector by cross-multiplying the first second unit vector in the second set of unit vectors and the second unit normal vector;

[0160] Step 950: Form a first basis vector matrix with the second first unit vector, the first unit normal vector, and the third unit vector in the first set of unit vectors, and form a second basis vector matrix with the second unit vector, the second unit normal vector, and the fourth unit vector in the second set of unit vectors;

[0161] Step 960: Calculate the rotation matrix of the sample base, which is obtained by multiplying the first basis vector matrix by the transpose matrix of the second basis vector matrix, so as to convert the first target point coordinates into the second target point coordinates by using the rotation matrix of the sample base.

[0162] The specific steps are as follows:

[0163] Select the base reference point, whose first reference point coordinates under the first microscopic imaging system are (x 14 , y 14 , z 14 ), and the second reference point coordinates under the second microscopic imaging system are (x 15 , y 15 , z 15 ).

[0164] If the base marking points include two linearly independent points, and their first base marking point coordinates are (x 16 , y 16 , z 16 ) and (x 17 , y 17 , z 17 ) respectively, calculate two first vectors v 11 = (x 16 - x 14 , y 16 - y 14 , z 16 - z 14 ), v 12 = (x 17 - x 14 , y 17 - y 14 , z 17 - z 14 ) as the first group of first vectors.

[0165] Under the second microscopic imaging system, the corresponding two second base marking point coordinates are (x 18 , y 18 , z 18 ) and (x 19 , y 19 , z 19 ) respectively, calculate two second vectors w 11 = (x 18 - x 15 , y 18 - y 15 , z 18 - z 15 ), w 12 = (x 19 - x 15 , y19 -y 15 , z 19 -z 15 ), as the second vector of the first group;

[0166] Calculate the first unit normal vector n1: Through the cross product of v 11 and v 12 , obtain n1 = v 11 × v 12 .

[0167] Normalize: n1 = n1 / |n1|.

[0168] Similarly, calculate the second unit normal vector n2: Through the cross product of w 11 and w 12 , obtain n2 = w 11 × w 12 , and after normalization, n2 = n2 / |n2|.

[0169] Select the second group of first vectors v 21 and v 22 that are linearly independent of n1, and normalize to obtain the first group of unit vectors u 11 = v 21 / | v 21| and u 12 = v 22 / | v 22| .

[0170] Select the second group of second vectors w 21 and w 22 that are linearly independent of n2, and normalize to obtain the second group of unit vectors u 21 = w 21 / | w 21| and u 22＝ w 22 / | w 22| .

[0171] Calculate the third unit vector u3 = n1 × u 11 , and after normalization, obtain u3 = u3 / | u 3| .

[0172] Calculate the fourth unit vector u4 = n2 × u 21 , and after normalization, obtain u4 = u4 / | u 4| .

[0173] Construct the first basis vector matrix B1 = [n1, u3, u 12 and the second basis vector matrix B2 = [n2, u4, u 22 .

[0174] Rotation matrix wherein is the transpose of B1.

[0175] Furthermore, the first target point coordinates (x1, y1) are transformed by the first basis vector matrix B1, the second basis vector matrix B2, and the rotation matrix R to obtain the second target point coordinates (x 18 , y 18 , z 18 ).

[0176] Use (x 18 , y 18 , z 18 ) to adjust the sample position.

[0177] As a way, as Figure 10 shown, use the rotation matrix of the sample base to convert the first target point coordinates into the second target point coordinates, which specifically includes the following steps:

[0178] Step 1010: Calculate the third vector of the first target point coordinates relative to the first reference point coordinates, and multiply the third vector by the first basis vector matrix to obtain the first basis vector representation;

[0179] Step 1020: Multiply the rotation matrix of the sample base by the first basis vector representation to obtain the fourth vector under the second microscopic imaging system;

[0180] Step 1030: Use the second basis vector matrix to convert the fourth vector into the second basis vector representation, and add the second reference point coordinates to the second basis vector representation to obtain the second target point coordinates.

[0181] In this embodiment, the rotation matrix R is constructed by the base reference point and multiple base marking points. The rotation matrix R is generated by the transformation of the basis vector matrix, reflecting the rotation relationship of the base in three-dimensional space. This embodiment refines the conversion process of the target point coordinates and realizes the mapping from the first microscopic imaging system to the second microscopic imaging system by using vector and matrix operations. The combination of the basis vector matrix and the rotation matrix ensures the accurate conversion of coordinates in three-dimensional space.

[0182] Specifically described as follows:

[0183] Calculate the third vector v3 = (x1 - x 14 , y1 - y 14 , z1 - z 14 ) for the first target point coordinates.

[0184] Project v3 onto the first basis vector matrix to obtain the first basis vector representation wherein is the inverse matrix of B1 (since B1 is an orthogonal matrix, ).

[0185] Calculate the fourth vector v4 = R·c1 using the rotation matrix R.

[0186] Obtain the second basis vector representation using the second basis vector matrix transformation (similarly, ).

[0187] The coordinates of the second target point (x4, y4, z4) = B2·c2+(x 15 , y 15 , z 15 ). Move the sample using the calculated coordinates of the second target point (x4, y4, z4) through the sample stage adjustment device.

[0188] This embodiment can counteract spatial vibration displacement.

[0189] As a further implementation, the marking points include the base marking points and the sample marking points, and the coordinates of the sample marking points under the first microscopic imaging system are the first marking point coordinates;

[0190] As Figure 11 shown, the conversion of the first target point coordinates into the second target point coordinates using the rotation matrix of the sample base includes but is not limited to the following steps:

[0191] Step 1110: Calculate the fifth vector of the first marking point coordinates relative to the first reference point coordinates, multiply the fifth vector by the first basis vector matrix to obtain the third basis vector representation, multiply the third basis vector representation by the rotation matrix of the sample base to obtain the sixth vector under the second microscopic imaging system, convert the sixth vector into the fourth basis vector representation using the second basis vector matrix, and add the second reference point coordinates to the second basis vector representation to obtain the second marking point coordinates under the second microscopic imaging system;

[0192] Step 1120: Calculate the seventh vector of the first target point coordinates relative to the first marking point coordinates, multiply the seventh vector by the first basis vector matrix to obtain the fifth basis vector representation, multiply the fifth basis vector representation by the rotation matrix of the sample base to obtain the eighth vector under the second microscopic imaging system, convert the eighth vector into the sixth basis vector representation using the second basis vector matrix, and add the second marking point coordinates to the second basis vector representation to obtain the second target point coordinates.

[0193] This embodiment describes a three-dimensional fine positioning algorithm, which corrects the position by introducing sample marker points on the basis of the preliminary positioning in the above embodiment. The sample marker points are located on the surface of the sample and are closer to the target point than the base marker points. The error caused by the base rotation matrix R is corrected by their relative positions, thereby improving the positioning accuracy. The correction process involves two vector conversions to determine the second target point coordinates of the sample marker points and the target point respectively.

[0194] The specific description is as follows:

[0195] The first marker point coordinates are (x7, y7, z7), and the fifth vector v5 = (x7 - x 14 , y7 - y 14 , z7 - z 14 ) is calculated.

[0196] The third base vector representation

[0197] The sixth vector v6 = R·c3.

[0198] The fourth base vector representation

[0199] The second marker point coordinates p m = B2·c4 + (x 15 , y 15 , z 15 ).

[0200] The seventh vector v7 = (x1 - x7, y1 - y7, z1 - z7) is calculated.

[0201] The fifth base vector representation

[0202] The eighth vector v8 = R·c5.

[0203] The sixth base vector representation

[0204] The second target point coordinates p2(x4, y4, z4) = p m + B2·c6.

[0205] In this embodiment, the fine positioning accuracy is improved. The spatial rotation error is reduced by correcting with the sample marker points, and the ability to resist thermal expansion interference is enhanced.

[0206] As another embodiment, the marker points include the base marker points and the sample marker points, and the coordinates of the sample marker points under the first microscopic imaging system are the first marker point coordinates;

[0207] As Figure 12 shown, converting the first target point coordinates into the second target point coordinates by using the rotation matrix of the sample base includes:

[0208] Step 1210: Calculate the ninth vector of the coordinates of the first marked point relative to the coordinates of the first reference point, multiply the ninth vector by the first basis vector matrix to obtain a fifth basis vector representation, multiply the fifth basis vector representation by the rotation matrix of the sample base to obtain a tenth vector under the second microscopic imaging system, convert the tenth vector into a sixth basis vector representation using the second basis vector matrix, and add the sixth basis vector representation to the coordinates of the second reference point to obtain the coordinates of the second marked point under the second microscopic imaging system;

[0209] Step 1220: Select a sample reference point on the sample surface of the sample, and obtain its first sample reference point coordinates under the first microscopic imaging system and its second sample reference point coordinates under the second microscopic imaging system;

[0210] Step 1230: Calculate the eleventh vector of the coordinates of the first sample marked point relative to the coordinates of the first sample reference point, and calculate the twelfth vector of the coordinates of the second base marked point relative to the coordinates of the second reference point;

[0211] Step 1240: Select a first group of eleventh vectors and a first group of twelfth vectors. The first group of eleventh vectors includes the eleventh vectors of two linearly independent base marked points, and the first group of twelfth vectors includes the twelfth vectors of the same base marked points corresponding to the eleventh vectors. Calculate a third unit normal vector based on the first group of eleventh vectors, and calculate a fourth unit normal vector based on the first group of twelfth vectors;

[0212] Step 1250: Select a second group of eleventh vectors that are linearly independent of the third unit normal vector and a second group of twelfth vectors that are linearly independent of the fourth unit normal vector and correspond to the same sample marked points as the second group of eleventh vectors. Normalize the second group of eleventh vectors to obtain a third group of unit vectors, and normalize the second group of twelfth vectors to obtain a fourth group of unit vectors. The third group of unit vectors includes two fifth unit vectors, and the fourth group of unit vectors includes two sixth unit vectors;

[0213] Step 1260: Perform a cross product of the first fifth unit vector in the third group of unit vectors and the third unit normal vector to calculate a seventh unit vector, and perform a cross product of the first sixth unit vector in the fourth group of unit vectors and the fourth unit normal vector to calculate an eighth unit vector;

[0214] Step 1270: Construct a third basis vector matrix from the second fifth unit vector, the third unit normal vector, and the seventh unit vector in the third set of unit vectors, and construct a fourth basis vector matrix from the sixth unit vector, the fourth unit normal vector, and the eighth unit vector in the fourth set of unit vectors;

[0215] Step 1280: Calculate the rotation matrix of the sample surface, which is obtained by multiplying the third basis vector matrix by the transpose matrix of the fourth basis vector matrix;

[0216] Step 1290: Calculate the thirteenth vector of the first target point coordinate relative to the first sample reference point coordinate, multiply the thirteenth vector by the third basis vector matrix to obtain a seventh basis vector representation, multiply the seventh basis vector representation by the rotation matrix of the sample surface to obtain a fourteenth vector under the second microscopic imaging system, convert the fourteenth vector into an eighth basis vector representation using the fourth basis vector matrix, and add the eighth basis vector representation to the second sample reference point coordinate to obtain the second target point coordinate.

[0217] This embodiment describes a three-dimensional fine positioning algorithm. By introducing a sample surface reference point based on the base rotation matrix, the rotation matrix R of the sample surface is calculated s . The sample surface reference point captures the independent rotation of the sample. Combining with the base rotation matrix R, the position of the target point is further corrected to improve the high-precision positioning ability.

[0218] Specifically described as follows:

[0219] Calculate the ninth vector v9 = (x7 - x 14 , y7 - y 14 , z7 - z 14 ).

[0220] Fifth basis vector representation

[0221] Tenth vector v 10 = R · c5.

[0222] Sixth basis vector representation

[0223] Second marker point coordinate p m = B2 · c6 + (x 15 , y 15 , z 15 ).

[0224] If the two first marker point coordinates are (x7, y7, z7) and (x 21 , y 21 , z 21), and the corresponding second marker point coordinates are (x 22 , y 22 , z 22 ) and (x 23 , y 23 , z 23 ).

[0225] Select the sample surface reference points. The coordinates of the first sample reference point are (x 24 , y 24 , z 24 ), and the coordinates of the second sample reference point are (x 25 , y 25 , z 25 ).

[0226] Calculate two eleventh vectors v 13 = (x7 - x 24 , y7 - y 24 , z7 - z 24 ) and v 14 = (x 21 - x 24 , y 21 - y 24 , z 21 - z 24 ) as the first group of eleventh vectors. Calculate two twelfth vectors w 23 = (x 22 - x 25 , y 22 - y 25 , z 22 - z 25 ) and w 24 = (x 23 - x 25 , y 23 - y 25 , z 23 - z 25 ) as the second group of twelfth vectors.

[0227] Calculate the third unit normal vector of the sample surface: n3 = v 13 × v 14 , and normalize n3 = n3 / |n3|.

[0228] Calculate the fourth unit normal vector of the sample surface: n4 = w 23 × w 24 , and normalize n4 = n4 / |n4|.

[0229] Select a second group of eleventh vectors v 15 and v 16 that are linearly independent of n3, and normalize them to obtain a third group of unit vectors, which includes two fifth unit vectors u 51= v 15 / | v 15 | and u 52 = v 16 / | v 16 |。

[0230] Select a second set of twelve vectors w that is linearly independent of n4 15 and w 16 , and normalize to obtain a fourth set of unit vectors, which contains two sixth unit vectors u 61 = w 15 / | w 15 | and u 62 = w 16 / | w 16 |。

[0231] Calculate the seventh unit vector u7 = n3 × u 51 , and after normalization, obtain u7 = u7 / |u7|.

[0232] Calculate the eighth unit vector u8 = n4 × u 61 , and after normalization, obtain u8 = u8 / |u8|.

[0233] Construct the third basis vector matrix B3 = [n3, u 52 , u7] and the fourth basis vector matrix B2 = [n4, u 62 , u8].

[0234] Rotation matrix where is the transpose of B1.

[0235] Sample plane rotation matrix

[0236] Calculate the thirteenth vector v 13 = (x1 - x 24 , y1 - y 24 , z1 - z 24 ).

[0237] Seventh basis vector representation

[0238] Fourteenth vector v 14 = R s lc7.

[0239] Eighth basis vector representation

[0240] The coordinates of the second target point p2 = B4 · c8 + (x 25 , y 25 , z 25 ).

[0241] Using this embodiment, the fine positioning accuracy is further improved, the complex spatial rotation error is reduced, and the vibration and thermal expansion interferences are counteracted.

[0242] An embodiment of the present disclosure also provides a sample positioning system. As Figure 13 shown, the sample positioning system 1300 is applicable to sample positioning across a first microscopic imaging system and a second microscopic imaging system, and includes:

[0243] A first acquisition module 1310, which acquires the first target point coordinates of a sample target point and the coordinates of at least one marker point under the first microscopic imaging system, where the marker point includes a base marker point and / or a sample marker point;

[0244] A second acquisition module 1320, which acquires the coordinates corresponding to the marker point under the second microscopic imaging system;

[0245] A calculation module 1330, which calculates the coordinate transformation relationship between the first microscopic imaging system and the second microscopic imaging system based on the coordinates of the marker point under the first microscopic imaging system and the second microscopic imaging system, and uses the coordinate transformation relationship to convert the first target point coordinates into the second target point coordinates under the second microscopic imaging system;

[0246] An adjustment module 1340, which adjusts the position of the sample in the second microscopic imaging system according to the second target point coordinates.

[0247] This system and device are composed of a first acquisition module 1310, a second acquisition module 1320, a calculation module 1330, and an adjustment module 1340. Each module can be interconnected through a data interface. The sample positioning system can be embedded into a front-end development tool as an independent software module, or can provide an interface as a cloud service for developers to remotely call.

[0248] This sample positioning system has a good anti-interference ability for the small position changes of the sample caused by physical properties (such as vibration, thermal expansion, etc.) through automatic acquisition and calculation of the coordinate transformation relationship. At the same time, it has a good processing effect on the physical position changes such as displacement, rotation, and tilt generated by the sample placement, can efficiently achieve the precise alignment of the sample target points between different microscopic imaging systems, avoid the errors introduced by traditional manual alignment, improve the experimental repeatability and data accuracy, improve the operation efficiency, and is applicable to a variety of microscopic imaging systems.

[0249] In this embodiment, the above-mentioned modules in the sample positioning system cooperate to implement the sample positioning method of the above embodiment. Specifically, for the method steps implemented by each module, please refer to the above-mentioned embodiment of the sample positioning method, and will not be elaborated here.

[0250] Those skilled in the art will readily conceive of other embodiments of the present disclosure after considering the specification and practicing the invention disclosed herein. The present disclosure is intended to cover any variations, uses, or adaptations of the present disclosure that follow the general principles of the present disclosure and include known common general knowledge or conventional technical means in the technical field not disclosed in the present disclosure. The specification and examples are only regarded as exemplary, and the true scope and spirit of the present disclosure are pointed out by the appended claims.

Claims

1. A sample positioning method, characterized in that, Applicable to sample positioning across a first microscopic imaging system and a second microscopic imaging system. The sample positioning method includes: Under the first microscopic imaging system, obtain the first target point coordinates of the sample target point and the coordinates of at least one marker point, where the marker point includes a base marker point and / or a sample marker point; Obtain the corresponding coordinates of the marker point under the second microscopic imaging system; Based on the coordinates of the marker point under the first microscopic imaging system and the second microscopic imaging system, calculate the coordinate transformation relationship between the first microscopic imaging system and the second microscopic imaging system, and use the coordinate transformation relationship to convert the first target point coordinates into the second target point coordinates under the second microscopic imaging system; Adjust the position of the sample in the second microscopic imaging system according to the second target point coordinates.

2. The sample positioning method according to claim 1, wherein Both the first target point coordinates and the coordinates of at least one marker point are two-dimensional coordinates; The marker point includes the base marker point of the sample base. The coordinates of the base marker point under the first microscopic imaging system are the first base marker point coordinates, and the coordinates under the second microscopic imaging system are the second base marker point coordinates; The following method is used to obtain the rotation angle of the sample base as the coordinate transformation relationship: Select a base reference point on the sample base, and obtain its first reference point coordinates under the first microscopic imaging system and the second reference point coordinates under the second microscopic imaging system; Based on the first relative coordinates between the first base marker point coordinates and the first reference point coordinates, and the second relative coordinates between the second base marker point coordinates and the second reference point coordinates, calculate the rotation angle of the sample base, so as to use the rotation angle of the sample base to convert the first target point coordinates into the second target point coordinates.

3. The sample positioning method according to claim 2, wherein The conversion of the first target point coordinates into the second target point coordinates using the rotation angle of the sample base includes: Calculate the third relative coordinates between the first target point coordinates and the first reference point coordinates, use the rotation angle of the sample base to convert the third relative coordinates into the fourth relative coordinates under the second microscopic imaging system, and calculate the second target point coordinates according to the fourth relative coordinates and the second reference point coordinates.

4. The sample positioning method according to claim 2, wherein The marker point includes the base marker point and the sample marker point. The coordinates of the sample marker point under the first microscopic imaging system are the first marker point coordinates; The conversion of the first target point coordinates into the second target point coordinates using the rotation angle of the sample base includes: Calculate the fifth relative coordinates between the first marker point coordinates and the first reference point coordinates, use the rotation angle of the sample base to convert the fifth relative coordinates into the sixth relative coordinates under the second microscopic imaging system, and calculate the second marker point coordinates of the sample marker point according to the sixth relative coordinates and the second reference point coordinates. Calculate the seventh relative coordinate between the first target point coordinate and the first marker point coordinate, convert the seventh relative coordinate into the eighth relative coordinate under the second microscopic imaging system by using the rotation angle of the sample base, and calculate the second target point coordinate according to the eighth relative coordinate and the second marker point coordinate.

5. The sample positioning method according to claim 2, wherein, The marker points include the base marker points and the sample marker points, and the coordinates of the sample marker points under the first microscopic imaging system are the first marker point coordinates; The conversion of the first target point coordinate into the second target point coordinate by using the rotation angle of the sample base includes: Calculate the fifth relative coordinate between the first marker point coordinate and the first reference point coordinate, convert the fifth relative coordinate into the sixth relative coordinate under the second microscopic imaging system by using the rotation angle of the sample base, and calculate the second marker point coordinate of the sample marker points according to the sixth relative coordinate and the second reference point coordinate; Select a sample reference point on the sample surface of the sample, and obtain its first sample reference point coordinate under the first microscopic imaging system and its second sample reference point coordinate under the second microscopic imaging system; Calculate the rotation angle of the sample surface based on the ninth relative coordinate between the first marker point coordinate and the first sample reference point coordinate, and the tenth relative coordinate between the second marker point coordinate and the second sample reference point coordinate; Calculate the eleventh relative coordinate between the first target point coordinate and the first sample reference point coordinate, convert the eleventh relative coordinate into the twelfth relative coordinate under the second microscopic imaging system by using the rotation angle of the sample surface, and calculate the second target point coordinate according to the twelfth relative coordinate and the second sample reference point coordinate.

6. The sample positioning method according to claim 2, wherein There are multiple base marker points; the base reference point is one of the multiple base marker points, or the base reference point is any point on the base outside the multiple base marker points.

7. The sample positioning method according to claim 1, wherein The first target point coordinate and the coordinates of at least one marker point are all three-dimensional coordinates; The marker points include the base marker points of the sample base, the coordinates of the base marker points under the first microscopic imaging system are the first base marker point coordinates, and the coordinates under the second microscopic imaging system are the second base marker point coordinates; Obtain the rotation matrix of the sample base as the coordinate conversion relationship by the following method: Select a base reference point on the sample base, and obtain its first reference point coordinate under the first microscopic imaging system and its second reference point coordinate under the second microscopic imaging system; Calculate the first vector of the first base marker point coordinate relative to the first reference point coordinate, and calculate the second vector of the second base marker point coordinate relative to the second reference point coordinate; Select the first group of first vectors and the first group of second vectors. The first group of first vectors includes the first vectors of two linearly independent base marker points, and the first group of second vectors includes the second vectors of the two linearly independent base marker points. Calculate the first unit normal vector based on the first group of first vectors, and calculate the second unit normal vector based on the first group of second vectors; Select the second group of first vectors that are linearly independent of the first unit normal vector and the second group of second vectors that are linearly independent of the second unit normal vector. Normalize the second group of first vectors to obtain the first group of unit vectors, and normalize the second group of second vectors to obtain the second group of unit vectors. The first group of unit vectors includes two first unit vectors, and the second group of unit vectors includes two second unit vectors; Perform a cross product of the first first unit vector in the first group of unit vectors and the first unit normal vector to calculate the third unit vector, and perform a cross product of the first second unit vector in the second group of unit vectors and the second unit normal vector to calculate the fourth unit vector; Construct the first basis vector matrix from the second first unit vector, the first unit normal vector, and the third unit vector in the first group of unit vectors, and construct the second basis vector matrix from the second unit vector, the second unit normal vector, and the fourth unit vector in the second group of unit vectors; Calculate the rotation matrix of the sample base, which is obtained by multiplying the first basis vector matrix by the transpose matrix of the second basis vector matrix, so as to convert the first target point coordinates into the second target point coordinates by using the rotation matrix of the sample base.

8. The sample positioning method according to claim 7, wherein The conversion of the first target point coordinates into the second target point coordinates by using the rotation matrix of the sample base includes: Calculate the third vector of the first target point coordinates relative to the first reference point coordinates, and multiply the third vector by the first basis vector matrix to obtain the first basis vector representation; Multiply the rotation matrix of the sample base by the first basis vector representation to obtain the fourth vector under the second microscopic imaging system; Convert the fourth vector into the second basis vector representation by using the second basis vector matrix, and add the second reference point coordinates to the second basis vector representation to obtain the second target point coordinates.

9. The sample positioning method according to claim 7, wherein The marker points include the base marker points and the sample marker points, and the coordinates of the sample marker points under the first microscopic imaging system are the first marker point coordinates; The conversion of the first target point coordinates into the second target point coordinates by using the rotation matrix of the sample base includes: Calculate the fifth vector of the coordinates of the first marked point relative to the coordinates of the first reference point, multiply the fifth vector by the first basis vector matrix to obtain a third basis vector representation, multiply the third basis vector representation by the rotation matrix of the sample base to obtain a sixth vector under the second microscopic imaging system, convert the sixth vector into a fourth basis vector representation using the second basis vector matrix, and add the second basis vector representation to the coordinates of the second reference point to obtain the coordinates of the second marked point under the second microscopic imaging system; Calculate the seventh vector of the coordinates of the first target point relative to the coordinates of the first marked point, multiply the seventh vector by the first basis vector matrix to obtain a fifth basis vector representation, multiply the fifth basis vector representation by the rotation matrix of the sample base to obtain an eighth vector under the second microscopic imaging system, convert the eighth vector into a sixth basis vector representation using the second basis vector matrix, and add the second basis vector representation to the coordinates of the second marked point to obtain the coordinates of the second target point.

10. The sample positioning method according to claim 7, characterized in that, The marked points include the base marked points and the sample marked points, and the coordinates of the sample marked points under the first microscopic imaging system are the coordinates of the first marked point; The conversion of the coordinates of the first target point into the coordinates of the second target point using the rotation matrix of the sample base includes: Calculate the ninth vector of the coordinates of the first marked point relative to the coordinates of the first reference point, multiply the ninth vector by the first basis vector matrix to obtain a fifth basis vector representation, multiply the fifth basis vector representation by the rotation matrix of the sample base to obtain a tenth vector under the second microscopic imaging system, convert the tenth vector into a sixth basis vector representation using the second basis vector matrix, and add the sixth basis vector representation to the coordinates of the second reference point to obtain the coordinates of the second marked point under the second microscopic imaging system; Select a sample reference point on the sample surface of the sample, and obtain its first sample reference point coordinates under the first microscopic imaging system and its second sample reference point coordinates under the second microscopic imaging system; Calculate the eleventh vector of the coordinates of the first sample marked point relative to the coordinates of the first sample reference point, and calculate the twelfth vector of the coordinates of the second base marked point relative to the coordinates of the second reference point; Select a first group of eleventh vectors and a first group of twelfth vectors. The first group of eleventh vectors includes the eleventh vectors of two linearly independent base marked points, and the first group of twelfth vectors includes the twelfth vectors corresponding to the same base marked points as the eleventh vectors. Calculate a third unit normal vector based on the first group of eleventh vectors, and calculate a fourth unit normal vector based on the first group of twelfth vectors; Select a second group of eleven vectors that are linearly independent of the third unit normal vector and a second group of twelve vectors that are linearly independent of the fourth unit normal vector and correspond to the same sample marker points as the second group of eleven vectors. Normalize the second group of eleven vectors to obtain a third group of unit vectors, and normalize the second group of twelve vectors to obtain a fourth group of unit vectors. The third group of unit vectors includes two fifth unit vectors, and the fourth group of unit vectors includes two sixth unit vectors; Perform a cross product calculation of a seventh unit vector based on the first fifth unit vector in the third group of unit vectors and the third unit normal vector, and perform a cross product calculation of an eighth unit vector based on the first sixth unit vector in the fourth group of unit vectors and the fourth unit normal vector; Form a third basis vector matrix with the second fifth unit vector, the third unit normal vector, and the seventh unit vector in the third group of unit vectors, and form a fourth basis vector matrix with the sixth unit vector, the fourth unit normal vector, and the eighth unit vector in the fourth group of unit vectors; Calculate the rotation matrix of the sample plane, which is obtained by multiplying the third basis vector matrix by the transpose matrix of the fourth basis vector matrix; Calculate a thirteenth vector of the first target point coordinates relative to the first sample reference point coordinates, multiply the thirteenth vector by the third basis vector matrix to obtain a seventh basis vector representation, multiply the sample plane rotation matrix by the seventh basis vector representation to obtain a fourteenth vector under the second microscopic imaging system, convert the fourteenth vector into an eighth basis vector representation using the fourth basis vector matrix, and add the eighth basis vector representation to the second sample reference point coordinates to obtain the second target point coordinates.

11. A sample positioning system, characterized in that, Applicable to sample positioning across the first microscopic imaging system and the second microscopic imaging system. The sample positioning system includes: A first acquisition module that, under the first microscopic imaging system, acquires the first target point coordinates of a sample target point and the coordinates of at least one marker point, where the marker points include base marker points and / or sample marker points; A second acquisition module that acquires the corresponding coordinates of the marker points under the second microscopic imaging system; A calculation module that, based on the coordinates of the marker points under the first microscopic imaging system and the second microscopic imaging system, calculates the coordinate transformation relationship between the first microscopic imaging system and the second microscopic imaging system, and uses the coordinate transformation relationship to convert the first target point coordinates into the second target point coordinates under the second microscopic imaging system; An adjustment module that adjusts the position of the sample in the second microscopic imaging system according to the second target point coordinates.

Images